In vivo multiphoton nanosurgery on cortical neurons

Leonardo Sacconi; Rod P. O'Connor; Audrius Jasaitis; Alessio Masi; Mario Buffelli; Francesco S. Pavone

doi:10.1117/1.2798723

1 September 2007 In vivo multiphoton nanosurgery on cortical neurons

Leonardo Sacconi, Rod P. O'Connor, Audrius Jasaitis, Alessio Masi, Mario Buffelli, Francesco S. Pavone

Author Affiliations +

Journal of Biomedical Optics, Vol. 12, Issue 5, 050502 (September 2007). https://doi.org/10.1117/1.2798723

Abstract

Two-photon microscopy has been used to perform high spatial resolution imaging of spine plasticity in the intact neocortex of living mice. Multiphoton absorption has also been used as a tool for the selective disruption of cellular structures in living cells and simple organisms. In this work, we exploit the spatial localization of multiphoton excitation to perform selective lesions on the neuronal processes of cortical neurons in living mice expressing fluorescent proteins. Neurons are irradiated with a focused, controlled dose of femtosecond laser energy delivered through cranial optical windows. The morphological consequences are then characterized with time lapse 3-D two-photon imaging over a period of minutes to days after the procedure. This methodology is applied to dissect single dendrites with submicrometric precision without causing any visible collateral damage to the surrounding neuronal structures. The spatial precision of this method is demonstrated by ablating individual dendritic spines, while sparing the adjacent spines and the structural integrity of the dendrite. The combination of multiphoton nanosurgery and in vivo imaging in mammals represents a promising tool for neurobiology and neuropharmacology research.

©(2007) Society of Photo-Optical Instrumentation Engineers (SPIE)

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Leonardo Sacconi, Rod P. O'Connor, Audrius Jasaitis, Alessio Masi, Mario Buffelli, and Francesco S. Pavone "In vivo multiphoton nanosurgery on cortical neurons," Journal of Biomedical Optics 12(5), 050502 (1 September 2007). https://doi.org/10.1117/1.2798723

Published: 1 September 2007

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KEYWORDS

Dendrites

Spine

Neurons

In vivo imaging

Fluorescent proteins

Absorption

Two photon excitation microscopy

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